Finding the partner: FRET and beyond.

Given the importance of aberrant protein-protein interactions (PPIs) in disease, the recent drug discovery focuses on targeting the altered PPIs to treat the disease. In this context, identifying the atypical PPIs underlying the disease is critical for the development of diagnostics and therapeutics. Various biochemical, biophysical, and genetic methods have been reported to study PPIs. Here, we are giving a short account of those techniques with more emphasis on Förster resonance energy transfer (FRET), which can be used to monitor macromolecular interactions in live cells. Besides the basics of FRET, we explain the modifications of its application, like Single molecule FRET (smFRET), Fluorescence Lifetime Imaging Microscopy-FRET (FLIM-FRET), and photoswitching FRET. While smFRET is extensively used for evaluating the biology of nucleic acids and also to develop diagnostics, FLIM-FRET is widely exploited to study the PPIs underlying neurological disorders and cancer. Photoswitching FRET is a relatively newer technique and it has tremendous potential to unravel the significance of different PPIs. Besides these modifications, there are several advancements in the field by introducing new fluorophores. Identification of lanthanide chelates, quantum dots, and other nanoparticle fluorophores has revolutionized the applications of FRET in diagnostics and basic biology. Yet, these methods can be employed to study the interactions of only two molecules. Since the majority of the PPIs are multimeric complexes, we still need to improve our technologies to study these interactions in live cells in real-time.

Immune Stimulation via Wounding Alters Chemical Profiles of Adult Tribolium castaneum.

Group-living individuals experience immense risk of disease transmission and parasite infection. In social and in some non-social insects, disease control with immunomodulation arises not only via individual immune defenses, but also via infochemicals such as contact cues and (defensive) volatiles to mount a group-level immunity. However, little is known about whether activation of the immune system elicits changes in chemical phenotypes, which may mediate these responses. We here asked whether individual immune experience resulting from wounding or injection of heat-killed Bacillus thuringiensis (priming) leads to changes in the chemical profiles of female and male adult red flour beetles, Tribolium castaneum, which are non-social but gregarious. We analyzed insect extracts using GC-FID to study the chemical composition of (1) cuticular hydrocarbons (CHCs) as candidates for the transfer of immunity-related information between individuals via contact, and (2) stink gland secretions, with analysis of benzoquinones as main active compounds regulating 'external immunity'. Despite a pronounced sexual dimorphism in CHC profiles, wounding stimulation led to similar profile changes in males and females with increases in the proportion of methyl-branched alkanes compared to naïve beetles. While changes in the overall secretion profiles were less pronounced, absolute amounts of benzoquinones were transiently elevated in wounded compared to naïve females. Responses to priming were insignificant in CHCs and secretions. We suggest that changes in different infochemicals after wounding may mediate immune status signaling in the context of both internal and external immune responses in groups of this non-social insect, thus showing parallels to social immunity.

Goat as the ideal climate-resilient animal model in tropical environment: revisiting advantages over other livestock species.

In the agriculture sector, livestock are considered extremely resilient to climate change and are tipped to play a significant role in ensuring food security to meet the increased demands of growing human population by 2050. Compared to other domestic species, goats are considered the ideal animal model for climate change due to its high thermal and drought resilience, ability to survive on limited pastures, and high disease resistance. This review is therefore a revisit to the advantages of rearing goats over other livestock species under current and future trends of changes in climate, particularly to cope with recurrent multiple stressors such as heat load, and lack of water and feed. In summary, goats, also called as poor man's cow, are preferred by the small-scale landless farmers due to their low input and assured higher output system, as they require low initial investment, with minimum specialized facilities and labors. Furthermore, they perceive goats as better resilient animal to cope with multiple stressors such as heat load, and water and feed scarcity, and possess better skills to cope with bush, when compared with sheep and cattle. The unique capacity for employing behavioral plasticity and morphological features of goats gives them clear advantage over sheep and cattle, when coping with seasonal biotopes, and experiences of water and feed shortage. When facing with low-quality feed, they also are superior to cattle and sheep to digest dry matter and to recycle nitrogen. Additionally, goats have superior ability to desiccate feces and concentrate urine, when compared with sheep and cattle. These advantages make goat the go-to species for efficiently countering the adversities associated with climate change and to optimize appropriate economic return through sustained production. Therefore, goats are tipped to be the future animals with extreme potential to counter the projected alarming climate change impacts and expected to play a significant role in ensuring food security to meet the demands of the growing human population by the end of this century.

Assessment of groundwater vulnerability to contamination: a case study.

Groundwater is a globally important, valuable resource for human life and economic development. Despite its importance, it is often misused and rarely well managed. Delineation of polluted zone attains significance especially when the pollution affects the human health. Delineation can be done by many approaches: process based, statistical and overlay and index methods. Overlay and index methods are relatively simpler and are based on hydrogeological settings and other factors which are considered to control the groundwater quality in a region. Groundwater vulnerability to contamination due to anthropogenic as well as seawater intrusion in an unconfined aquifer spread over the watershed areas of Kazhakkoottam, Kulathoor and Menamkulam regions of Thiruvananthapuram District, Kerala, India, was carried out using both susceptibility index (SI) method and modified GALDIT index method. Majority of the region selected was categorised as moderately vulnerable by both indices. The cause of groundwater vulnerability to contamination was found using factor analysis. The factor score map of the study area also shows that majority of the study area is moderately affected by contamination. Hence, the area requires continuous monitoring of groundwater quality to prevent the contamination of groundwater resources for future generations.

Ayurveda in the integrative management of bovine secondary recumbency-a case report.

Downer cow syndrome, or secondary recumbency, is a condition primarily affecting dairy cows, where the animal is unable to rise and stand, due to unknown cause. It is usually associated with poor prognosis. Terminal downers are euthanized in most countries. A four-year old Kasarkode dwarf-cow, post-calving was brought up laterally recumbent with heavy nasal discharge, labored breathing, loss of appetite and signs of dehydration on 6th day of its recumbency. Before this, the patient was diagnosed with milk fever and standard treatment with calcium borogluconate was administered intravenously. The animal was left to succumb under unprotected conditions, due to various constraints on euthanasia. After adopting the cow, Nasya was started immediately to avoid death due to sepsis and shock. The animal was drenched with Ayurvedic fluids containing deepana-pacana herbs. Sternal recumbency, warm and moistened muzzle was observed on the fourth day of commencing ayurvedic treatment. Respiratory distress was minimal. Drastic prognostic shift from "no hope" to "good" was possible within 6 days thanks to Nasya, and the animal was stable. Thereafter, integrative care comprising of antibiotics, rehydrating IV fluids, and supplementations, along with ayurvedic medicines was initiated. Ruminal-fluid obtained from slaughterhouse was used for ruminal-flora replacement. Rumination on 14th day, cow on its feet by 19th day and complete healing of decubital ulcers by approximately 40 days was recorded. A downed cow which did not respond to standard veterinary care was managed with Ayurveda-integrated veterinary care. Ayurveda herbs like bamboo leaves (Bambusa vulgaris), green chiretta (Andrographis paniculata) that cattle prefer eating during certain illness, turn out to be useful for Ayurvedic management. Hence, Ayurveda veterinary medicine might be, a good choice for integrative management of terminal downers, preventing early death in downed dairy cows.

Deciphering a Beetle Clock: Individual and Sex-Dependent Variation in Daily Activity Patterns.

Circadian clocks are inherent to most organisms, including cryptozoic animals that seldom encounter direct light, and regulate their daily activity cycles. A conserved suite of clock genes underpins these rhythms. In this study, we explore the circadian behaviors of the red flour beetle Tribolium castaneum, a significant pest impacting stored grain globally. We report on how daily light and temperature cues synchronize distinct activity patterns in these beetles, characterized by reduced morning activity and increased evening activity, anticipating the respective environmental transitions. Although less robust, rhythmicity in locomotor activity is maintained in constant dark and constant light conditions. Notably, we observed more robust rhythmic behaviors in males than females with individual variation exceeding those previously reported for other insect species. RNA interference targeting the Clock gene weakened locomotor activity rhythms. Our findings demonstrate the existence of a circadian clock and of clock-controlled behaviors in T. castaneum. Furthermore, they highlight substantial individual differences in circadian activity, laying the groundwork for future research on the relevance of individual variation in circadian rhythms in an ecological and evolutionary context.

Furan based synthetic chalcone derivative functions against gut inflammation and oxidative stress demonstrated in in-vivo zebrafish model.

Inflammatory Bowel Disease (IBD) is a group of persistent intestinal illnesses resulting from bowel inflammation unrelated to infection. The prevalence of IBD is rising in industrialized countries, increasing healthcare costs. Whether naturally occurring or synthetic, chalcones possess a broad range of biological properties, including anti-inflammatory, anti-microbial, and antioxidant effects. This investigation focuses on DKO7 (E)-3-(4-(dimethylamino)phenyl)-1-(5-methylfuran-2-yl)prop-2-en-1-one, a synthesized chalcone with potential anti-inflammatory effects in a zebrafish model of intestinal inflammation induced by Dextran sodium sulfate (DSS). The in vitro study displayed dose-dependent anti-inflammatory as well as antioxidant properties of DKO7. Additionally, DKO7 protected zebrafish larvae against lipid peroxidation, reactive oxygen stress (ROS), and DSS-induced inflammation. Moreover, DKO7 reduced the expression of pro-inflammatory genes, including TNF-α, IL-1ß, IL-6, and iNOS. Further, it reduced the levels of nitric oxide (NO) and lactate dehydrogenase (LDH) in the intestinal tissues of adult zebrafish and increased the levels of antioxidant enzymes such as Catalase (CAT) and superoxide dismutase (SOD). The protective effect of DKO7 against chemically (or DSS) induced intestinal inflammation was further verified using histopathological techniques in intestinal tissues. The furan-based chalcone derivative, DKO7, displayed antioxidant and anti-inflammatory properties. Also, DKO7 successfully reverses the DSS-induced intestinal damage in zebrafish. Overall, this study indicates the ability of DKO7 to alleviate DSS-induced gut inflammation in an in-vivo zebrafish.

Survival of the Sawfly Athalia rosae Upon Infection by an Entomopathogenic Fungus and in Relation to Clerodanoid Uptake.

Larvae of the turnip sawfly Athalia rosae are a pest of Brassicacae plants, as their feeding can cause defoliation of various crops of economic importance. The larvae and the adults of this sawfly species are known to take up different classes of chemical compounds from their respective host plants, with potentially deterrent functions against predators. In addition, compounds taken up by the adults, the clerodanoids, are known for their antimicrobial activity. These features could be a challenge to biocontrol strategies. Several natural enemies of A. rosae have been identified, targeting larval and pupal stages of A. rosae, which could potentially be used as biocontrol agents. However, targeting the adult stage of a larval pest in addition to targeting the juvenile stages may improve population control. In this study, we ask whether a strain of the entomopathogenic fungus Beauveria bassiana shows biological activity against A. rosae adults. We also investigate whether the behavior of clerodanoid uptake by the adults, which is commonly found, affects their survival in response to a B. bassiana exposure. We found a clear dose-response relationship, i.e., with increasing fungal conidia concentrations survival of A. rosae decreased. However, there was only a low incidence of mycelial growth and sporulation from A. rosae cadavers, indicating that either the fungus is not successfully developing inside this host, or it is not able to re-emerge from it. Clerodanoid uptake decreased the survival of healthy adults; however, it did not increase their survival to B. bassiana. Our results revealed that this strain of B. bassiana if applied alone is probably not suitable for biocontrol of this sawfly species, because A. rosae showed a high baseline resistance against this fungus. The behavior of clerodanoid uptake is unlikely to have evolved as a defense against this entomopathogenic fungus.

Reporters of Cancer Stem Cells as a Tool for Drug Discovery.

In view of the importance of cancer stem cells (CSCs) in chemoresistance, metastasis and recurrence, the biology of CSCs were explored in detail. Based on that, several modalities were proposed to target them. In spite of the several clinical trials, a successful CSC-targeting drug is yet to be identified. The number of molecules screened and entered for clinical trial for CSC-targeting is comparatively low, compared to other drugs. The bottle neck is the lack of a high-throughput adaptable screening strategy for CSCs. This review is aimed to identify suitable reporters for CSCs that can be used to identify the heterogeneous CSC populations, including quiescent CSCs, proliferative CSCs, drug resistant CSCs and metastatic CSCs. Analysis of the tumor microenvironment regulating CSCs revealed that the factors in CSC-niche activates effector molecules that function as CSC markers, including pluripotency markers, CD133, ABCG2 and ALDH1A1. Among these factors OCT4, SOX2, NANOG, ABCG2 and ALDH1A1 are ideal for making reporters for CSCs. The pluripotency molecules, like OCT4, SOX2 and NANOG, regulate self-renewal, chemoresistance and metastasis. ABCG2 is a known regulator of drug resistance while ALDH1A1 modulates self-renewal, chemoresistance and metastasis. Considering the heterogeneity of CSCs, including a quiescent population and a proliferative population with metastatic ability, we propose the use of a combination of reporters. A dual reporter consisting of a pluripotency marker and a marker like ALDH1A1 will be useful in screening drugs that target CSCs.

Electronic and Vibrational Decoupling in Chemically Exfoliated Bilayer Thin Two-Dimensional V2O5.

The synthesis of high-quality two-dimensional (2D) transition metal oxides is challenging compared to 2D transition metal dichalcogenides as a result of the exotic surface changes that can appear during formation. Herein, we report the synthesis of bilayer 2D V2O5 nanosheets with a thickness of ∼1 nm using the chemical exfoliation method and a comprehensive study on the vibrational and optical properties of bilayer 2D V2O5. We report, for the first time, a thickness-dependent blue shift of 1.33 eV in the optical bandgap, which signifies the emergence of electronic decoupling in bilayer 2D V2O5. In addition, a thickness-dependent vibrational decoupling of phonon modes observed via Raman spectroscopy fingerprinting was verified by computing the lattice vibrational modes using the density functional perturbation theory. We demonstrate that the manifestation of the electronic and vibrational decoupling can be used as a benchmark to confirm the successful formation of bilayer 2D V2O5 from its bulk counterpart.